Rapid detection of Staphylococcus aureus in food safety using an RPA-CRISPR-Cas12a assay

Lin, Liyun; Zha, Guangcai; Wei, Huagui; Zheng, Yuzhong; Yang, Ping; Lin, Min; Liu, Mouquan; Wang, Zhonghe; Zou, Xianghui; Zhu, Hai‐Liang; Luo, Qiulan; Li, Jinquan; Lin, Min

doi:10.1016/j.foodcont.2022.109505

Cited by 14 publications

(2 citation statements)

References 22 publications

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“…Moreover, they achieved the detection of three transgenes via the multiplexed microfluidic device embedded in LFD. Similar work based on RPA and Cas12a has also been used to detect HPV, 97,98 Staphylococcus aureus, 99 and drug-resistant genes. 100,101…”

Section: Crispr/cas System In Optical Biosensorsmentioning

confidence: 99%

Recent progress on the CRISPR/Cas system in optical biosensors

Li,

Zhai,

Dong

et al. 2024

Anal. Methods

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Section: Crispr/cas System In Optical Biosensorsmentioning

confidence: 99%

Recent progress on the CRISPR/Cas system in optical biosensors

Li,

Zhai,

Dong

et al. 2024

Anal. Methods

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“…Staphylococcus aureus [1], one of the top five foodborne pathogens, has a common and strong aggressiveness in humans and can secrete multiple toxic proteins (Pathogenic enterotoxins, Hemolysin, PVL) [1,2] which can cause bacteraemia, endocarditis, meningitis, toxic shock syndrome, pneumonia and other dangerous infectious diseases [3]. Moreover, the worldwide annual incidence rate of bacteremia is 15-40 cases per 100,000 people [4].…”

Section: Introductionmentioning

confidence: 99%

Isothermal Amplification and CRISPR/Cas12a-System-Based Assay for Rapid, Sensitive and Visual Detection of Staphylococcus aureus

Xu,

Zeng,

et al. 2023

Foods

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Staphylococcus aureus exists widely in the natural environment and is one of the main food-borne pathogenic microorganisms causing human bacteremia. For safe food management, a rapid, high-specificity, sensitive method for the detection of S. aureus should be developed. In this study, a platform for detecting S. aureus (nuc gene) based on isothermal amplification (loop-mediated isothermal amplification—LAMP, recombinase polymerase amplification—RPA) and the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas12a) proteins system (LAMP, RPA-CRISPR/Cas12a) was proposed. In this study, the LAMP, RPA-CRISPR/Cas12a detection platform and immunochromatographic test strip (ICS) were combined to achieve a low-cost, simple and visualized detection of S. aureus. The limit of visual detection was 57.8 fg/µL of nuc DNA and 6.7 × 102 CFU/mL of bacteria. Moreover, the platform could be combined with fluorescence detection, namely LAMP, RPA-CRISPR/Cas12a-flu, to establish a rapid and highly sensitive method for the detection of S. aureus. The limit of fluorescence detection was 5.78 fg/µL of genomic DNA and 67 CFU/mL of S. aureus. In addition, this detection platform can detect S. aureus in dairy products, and the detection time was ~40 min. Consequently, the isothermal amplification CRISPR/Cas12a platform is a useful tool for the rapid and sensitive detection of S. aureus in food.

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A Lateral Flow Assay Based on Streptavidin‐biotin Amplification System with Recombinase Polymerase Amplification for Rapid and Quantitative Detection of Salmonella enteritidis

Feng,

Fu,

Cao

et al. 2023

ChemBioChem

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Salmonella constitutes a prevalent alimentary pathogen, instigating zoonotic afflictions. Consequently, the prompt discernment of Salmonella in sustenance is of cardinal significance. Lateral flow assays utilizing colorimetric methodologies adequately fulfill the prerequisites of point‐of‐care diagnostics, however, their detection threshold remains elevated, generally permitting only qualitative discernment, an impediment to the preliminary screening of nascent pathogens. In response to this conundrum, we propose a lateral flow diagnostic predicated upon a streptavidin‐biotin amplification system with recombinase polymerase amplification engineered for the expeditious and quantitative discernment of Salmonella enteritidis. Trace nucleic acids within a sample undergo exponential amplification via recombinase polymerase amplification to a level discernable, constituting the initial signal amplification. Subsequently, along the test line (T‐line) of the lateral flow strip, the chromatic signal undergoes augmentation by securing a greater quantity of AuNPs through the magnification capacity of the streptavidin‐biotin mechanism, affecting the second signal amplification. Quantitative results are procured via smartphone capture and transferred to computer software for precise calculation of the targeted quantity. The lateral flow strip exhibits a LOD at 19.41 CFU/mL for cultured S. enteritidis. The RSD of three varying concentrations were respectively 3.74%, 5.96%, and 4.25%.

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Rapid detection of Staphylococcus aureus in food safety using an RPA-CRISPR-Cas12a assay

Cited by 14 publications

References 22 publications

Recent progress on the CRISPR/Cas system in optical biosensors

Recent progress on the CRISPR/Cas system in optical biosensors

Isothermal Amplification and CRISPR/Cas12a-System-Based Assay for Rapid, Sensitive and Visual Detection of Staphylococcus aureus

A Lateral Flow Assay Based on Streptavidin‐biotin Amplification System with Recombinase Polymerase Amplification for Rapid and Quantitative Detection of Salmonella enteritidis

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